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A fast-screening dispersive liquid–liquid microextraction–gas chromatography–mass spectrometry method applied to the determination of efavirenz in human plasma samples

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Abstract

We demonstrate the suitability of a fast, green, easy-to-perform, and modified sample extraction procedure, i.e., dispersive liquid–liquid microextraction (DLLME) for the determination of efavirenz (EFV) in human plasma. Data acquisition was done by gas chromatography–mass spectrometry (GC–MS) in the selected ion monitoring (SIM) mode. The simplicity of the method lies in, among others, the avoidance of the use of large organic solvent volumes as mobile phases and non-volatile buffers that tend to block the plumbing in high-performance liquid chromatography (HPLC). Chromatographic and mass spectral parameters were optimized using bovine whole blood for matrix matching due to insufficient human plasma. Method validation was accomplished using the United States Food and Drug Administration (USFDA) 2018 guidelines. The calibration curve was linear with a dynamic range of 0.10–2.0 μg/mL and an R2 value of 0.9998. The within-run accuracy and precision were both less than 20% at the lower limit of quantification (LLOQ) spike level. The LLOQ was 0.027 μg/mL which compared well with some values but was also orders of magnitude better than others reported in the literature. The percent recovery was 91.5% at the LLOQ spike level. The DLLME technique was applied in human plasma samples from patients who were on treatment with EFV. The human plasma samples gave concentrations of EFV ranging between 0.14–1.00 μg/mL with three samples out of seven showing concentrations that fell within or close to the recommended therapeutic range.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors thank the University of Botswana and the Ministry of Health and Wellness for granting ethical approval.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Department of Chemistry, Faculty of Science, University of Botswana, PB 00704, Gaborone, Botswana

    Wangu Masenga & Kwenga Sichilongo

  2. Botswana – University of Pennsylvania Partnership (BUP), Box AC 157 ACH, Gaborone, Botswana

    Giacomo Maria Paganotti

  3. Division of Infectious Diseases, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Giacomo Maria Paganotti

  4. Department of Biomedical Sciences, Faculty of Medicine, University of Botswana, PB 00713, Gaborone, Botswana

    Giacomo Maria Paganotti

  5. Botswana Harvard AIDS institute partnership (BHP), P.O. Box BO, 320, Gaborone, Botswana

    Kaelo Seatla & Simani Gaseitsiwe

  6. Department of Medical Laboratory Sciences, School of Allied Health Professionals, University of Botswana, Gaborone, Botswana

    Kaelo Seatla

  7. Department of Immunology & Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA, 02115, USA

    Simani Gaseitsiwe

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  1. Wangu Masenga
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Correspondence to Kwenga Sichilongo.

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Ethical approval to conduct this study was obtained from the University of Botswana Office of Research and Development (Reference Number RES/IRB/CHMS/014) and Human Research Development Division (HRDD) (Reference Number HPDME: 13/18/1).

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Masenga, W., Paganotti, G.M., Seatla, K. et al. A fast-screening dispersive liquid–liquid microextraction–gas chromatography–mass spectrometry method applied to the determination of efavirenz in human plasma samples. Anal Bioanal Chem 413, 6401–6412 (2021). https://doi.org/10.1007/s00216-021-03604-0

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